Mouse, Rat, Rabbit, Human, African green monkey
The pericentrin clone used is 1.7 kb in size and is derived from within residues 100-600 of mouse pericentrin 1. It was expressed as a fusion protein. The corresponding amino acids are present in both pericentrin and kendrin (pericentrin-2) so this antibody is predicted to cross-react with both isoforms.
ICC/IF: MCF7 and NIH3T3 cells.
存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
存储溶液pH: 7.40 Preservative: 0.02% Sodium azide Constituent: PBS Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 0.1 - 0.5 µg/ml.
Use at an assay dependent concentration.
Use at an assay dependent concentration.
功能Integral component of the filamentous matrix of the centrosome involved in the initial establishment of organized microtubule arrays in both mitosis and meiosis. Plays a role, together with DISC1, in the microtubule network formation. Is an integral component of the pericentriolar material (PCM). May play an important role in preventing premature centrosome splitting during interphase by inhibiting NEK2 kinase activity at the centrosome.
组织特异性Expressed in all tissues tested, including placenta, liver, kidney and thymus.
结构域Composed of a coiled-coil central region flanked by non-helical N- and C-terminals.
细胞定位Cytoplasm > cytoskeleton > microtubule organizing center > centrosome. Centrosomal at all stages of the cell cycle. Remains associated with centrosomes following microtubule depolymerization. Colocalized with DISC1 at the centrosome.
ICC/IF image of ab4448 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab4448, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
ab4448 stained NIH-3T3 cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab4448 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Immunocytochemistry/ Immunofluorescence - Anti-Pericentrin antibody - Centrosome Marker (ab4448)This image is courtesy of Roberto Giambruno, Marilena Ciciarello and Patrizia Lavia
NIH3T3 cells were fixed in 100% methanol for 6 minutes at -20°C, washed 3 times in PBS then incubated with ab4448 (1/2000) for 1 hour at room temperature. The panel of images shows the nuclei stained with DAPI (blue), ab4448 staining is shown in green. 100x magnification.
Paraffin embedded human kidney tissue sections were incubated with ab4448 (1/4500 dilution) for 30 minutes at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.
ab4448 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further images can be found at www.proteinatlas.org
Immunocytochemistry/ Immunofluorescence - Anti-Pericentrin antibody - Centrosome Marker (ab4448)This image is courtesy of Gordon Chan, University of Alberta
IF staining of pericentrin in MCF7 cells (human).
The top panel is an interphase cell showing centrosome staining. The bottom panel shows a mitotic cell with spindle pole staining. ab4448 was used at 1/500, but also works at higher dilutions (1/1000-1/2000).
Top panel - 630X magnification; Bottom panel -1000X magnification. The secondary antibody was Alexa488 anti-rabbit.
ICC/IF image of ab4448 stained Hela cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab4448, 1µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Subramanian L et al. Dynamic behaviour of human neuroepithelial cells in the developing forebrain. Nat Commun8:14167 (2017).
Read more (PubMed: 28139695) »
Rickardson L et al. Evaluation of the antitumor activity of NOV202, a novel microtubule targeting and vascular disrupting agent. Drug Des Devel Ther11:1335-1351 (2017).
Read more (PubMed: 28496304) »